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Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm

(1) Background: Whole Exome Sequencing of patients with thoracic aortic aneurysm often identifies “Variants of Uncertain Significance” (VUS), leading to uncertainty in clinical management. We assess a novel mechanism for potential routine assessment of these genes in TAA patients. Zebrafish are incr...

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Autores principales: Prendergast, Andrew, Ziganshin, Bulat A., Papanikolaou, Dimitra, Zafar, Mohammad A., Nicoli, Stefania, Mukherjee, Sandip, Elefteriades, John A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8775119/
https://www.ncbi.nlm.nih.gov/pubmed/35052463
http://dx.doi.org/10.3390/genes13010123
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author Prendergast, Andrew
Ziganshin, Bulat A.
Papanikolaou, Dimitra
Zafar, Mohammad A.
Nicoli, Stefania
Mukherjee, Sandip
Elefteriades, John A.
author_facet Prendergast, Andrew
Ziganshin, Bulat A.
Papanikolaou, Dimitra
Zafar, Mohammad A.
Nicoli, Stefania
Mukherjee, Sandip
Elefteriades, John A.
author_sort Prendergast, Andrew
collection PubMed
description (1) Background: Whole Exome Sequencing of patients with thoracic aortic aneurysm often identifies “Variants of Uncertain Significance” (VUS), leading to uncertainty in clinical management. We assess a novel mechanism for potential routine assessment of these genes in TAA patients. Zebrafish are increasingly used as experimental models of disease. Advantages include low cost, rapid maturation, and physical transparency, permitting direct microscopic assessment. (2) Methods: Zebrafish loss of function mutations were generated using a CRISPRC/CAS9 approach for EMILIN1 and MIB1 genes similar to VUSs identified in clinical testing. Additionally, “positive control” mutants were constructed for known deleterious variants in FBN1 (Marfan’s) and COL1A2, COL5A1, COL5A2 (Ehlers-Danlos). Zebrafish embryos were followed to six days post-fertilization. Embryos were studied by brightfield and confocal microscopy to ascertain any vascular, cardiac, and skeletal abnormalities. (3) Results: A dramatic pattern of cardiac, cerebral, aortic, and skeletal abnormalities was identified for the known pathogenic FBN1 and COL1A2, COL5A1, and COL5A2 mutants, as well as for the EMILIN1 and MIB1 mutants of prior unknown significance. Visualized abnormalities included hemorrhage (peri-aortic and cranial), cardiomegaly, reduced diameter of the aorta and intersegmental vessels, lower aortic cell counts, and scoliosis (often extremely severe). (4) Conclusion: This pilot study suggests that candidate genes arising in clinical practice may be rapidly assessed via zebrafish mutants—thus permitting evidence-based decisions about pathogenicity. Thus, years-long delays to clinically demonstrate pathogenicity may be obviated. Zebrafish data would represent only one segment of analysis, which would also include frequency of the variant in the general population, in silico genetic analysis, and degree of preservation in phylogeny.
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spelling pubmed-87751192022-01-21 Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm Prendergast, Andrew Ziganshin, Bulat A. Papanikolaou, Dimitra Zafar, Mohammad A. Nicoli, Stefania Mukherjee, Sandip Elefteriades, John A. Genes (Basel) Article (1) Background: Whole Exome Sequencing of patients with thoracic aortic aneurysm often identifies “Variants of Uncertain Significance” (VUS), leading to uncertainty in clinical management. We assess a novel mechanism for potential routine assessment of these genes in TAA patients. Zebrafish are increasingly used as experimental models of disease. Advantages include low cost, rapid maturation, and physical transparency, permitting direct microscopic assessment. (2) Methods: Zebrafish loss of function mutations were generated using a CRISPRC/CAS9 approach for EMILIN1 and MIB1 genes similar to VUSs identified in clinical testing. Additionally, “positive control” mutants were constructed for known deleterious variants in FBN1 (Marfan’s) and COL1A2, COL5A1, COL5A2 (Ehlers-Danlos). Zebrafish embryos were followed to six days post-fertilization. Embryos were studied by brightfield and confocal microscopy to ascertain any vascular, cardiac, and skeletal abnormalities. (3) Results: A dramatic pattern of cardiac, cerebral, aortic, and skeletal abnormalities was identified for the known pathogenic FBN1 and COL1A2, COL5A1, and COL5A2 mutants, as well as for the EMILIN1 and MIB1 mutants of prior unknown significance. Visualized abnormalities included hemorrhage (peri-aortic and cranial), cardiomegaly, reduced diameter of the aorta and intersegmental vessels, lower aortic cell counts, and scoliosis (often extremely severe). (4) Conclusion: This pilot study suggests that candidate genes arising in clinical practice may be rapidly assessed via zebrafish mutants—thus permitting evidence-based decisions about pathogenicity. Thus, years-long delays to clinically demonstrate pathogenicity may be obviated. Zebrafish data would represent only one segment of analysis, which would also include frequency of the variant in the general population, in silico genetic analysis, and degree of preservation in phylogeny. MDPI 2022-01-10 /pmc/articles/PMC8775119/ /pubmed/35052463 http://dx.doi.org/10.3390/genes13010123 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Prendergast, Andrew
Ziganshin, Bulat A.
Papanikolaou, Dimitra
Zafar, Mohammad A.
Nicoli, Stefania
Mukherjee, Sandip
Elefteriades, John A.
Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title_full Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title_fullStr Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title_full_unstemmed Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title_short Phenotyping Zebrafish Mutant Models to Assess Candidate Genes Associated with Aortic Aneurysm
title_sort phenotyping zebrafish mutant models to assess candidate genes associated with aortic aneurysm
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8775119/
https://www.ncbi.nlm.nih.gov/pubmed/35052463
http://dx.doi.org/10.3390/genes13010123
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